Transactions of the Korean hydrogen and new energy society (한국수소및신에너지학회논문집)

The Korean Hydrogen and New Energy Society (한국수소및신에너지학회)
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Purification of Hydrogenase from Thiocapsa roseopersicina : Effect of Ammonium Sulfate Precipitation and Heat-Treatment

홍색 유황세균 Thiocapsa roseopersicina가 생산하는 수소생산 효소의 정제 : Ammonium sulfate 침전 및 열처리의 효과

  • Choi, Eun-Hye (Bioenergy Research Center, Korea Institute of Energy Research) ;
  • Oh, You-Kwan (Bioenergy Research Center, Korea Institute of Energy Research) ;
  • Kim, Mi-Sun (Bioenergy Research Center, Korea Institute of Energy Research)
  • 최은혜 (한국에너지기술연구원 바이오에너지연구센터) ;
  • 오유관 (한국에너지기술연구원 바이오에너지연구센터) ;
  • 김미선 (한국에너지기술연구원 바이오에너지연구센터)
  • Published : 2006.12.15
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Abstract

Effect of $(NH_4)_2SO_4$ precipitation and heat-treatment on hydrogenase which was extracted from the cytoplasmic fraction of the phototrophic purple sulfur bacterium Thiocapsa roseopersicina NCIB 8347 was studied. Crude enzyme extract was prepared by centrifugation($28,000{\times}g$, $400,000{\times}g$) after sonication of cells grown under photosynthetic condition for 96 hrs. Various conditions of $(NH_4)_2SO_4$ precipitation and heat-treatment were examined and the effect of protein concentration was analyzed by SDS-electrophoresis between the treatments. Optimum conditions for $(NH_4)_2SO_4$ precipitation and heat-treatment for evolution hydrogenase activity were 40-60% saturation and $60^{\circ}C$ for 20 min, respectively, which exhibited the specific hydrogenase activity of 0.78 U/mg-protein. Specific hydrogenase activity was decreased to 31.6% when the heat-treatment at $60^{\circ}C$ increased from 20 min to 5 hrs.

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References

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